Protein synthesis is a fundamental biological process that underlies the development of polypeptide therapeutics, vaccines, diagnostics, and industrial enzymes. With the advent of recombinant DNA (rDNA) technology, it has become possible to harness the catalytic machinery of the cell to produce a desired protein. This can be achieved within the cellular environment or in vitro using extracts derived from cells.
Methods of recombinant protein synthesis include cell-free and cell-based methods of protein synthesis to introduce nonnatural amino acids into a protein. For example, in vitro translation has been recognized for its ability to incorporate nonnatural and isotope-labeled amino acids as well as its capability to produce proteins that are unstable, insoluble, or cytotoxic in vivo. In addition, cell-free protein synthesis may play a role in revolutionizing protein engineering and proteomic screening technologies. The cell-free method bypasses the laborious processes required for cloning and transforming cells for the expression of new gene products in vivo, and is becoming a platform technology for this field.
Relevant Literature
In vivo production of a green fluorescent protein containing multiple copies of pAzF is discussed in Johnson et al. (2011) Nature Chemical Biology 7:779-786. U.S. Pat. Nos. 7,375,234; 7,718,410; 8,030,074; 7,045,337. U.S. Patent Application 20080311412.
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